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Small Molecule Screening of Primary Human Acute Myeloid Leukemia Using Co-culture and Multiplexed FACS Analysis

Baudet, Aurélie LU ; Hultmark, Simon LU ; Ek, Fredrik LU and Magnusson, Mattias LU (2022) In Bio-protocol 12(6).
Abstract

Ex vivo culture of primary acute myeloid leukemia (AML) cells is notoriously difficult due to spontaneous differentiation and cell death, which hinders mechanistic and translational studies. To overcome this bottleneck, we have implemented a co-culture system, where the OP9-M2 stromal cells support the growth, but most notably limit the differentiation of primary AML cells, thus allowing for mechanistic studies in vitro. Additionally, the co-culture on OP9-M2 stromal is superior in preserving surface marker expression of primary (adult and pediatric) AML cells in comparison to stroma-free culture. Thus, by combining the co-culture with multicolor, high-throughput FACS, we can evaluate the effect of hundreds of small molecules on... (More)

Ex vivo culture of primary acute myeloid leukemia (AML) cells is notoriously difficult due to spontaneous differentiation and cell death, which hinders mechanistic and translational studies. To overcome this bottleneck, we have implemented a co-culture system, where the OP9-M2 stromal cells support the growth, but most notably limit the differentiation of primary AML cells, thus allowing for mechanistic studies in vitro. Additionally, the co-culture on OP9-M2 stromal is superior in preserving surface marker expression of primary (adult and pediatric) AML cells in comparison to stroma-free culture. Thus, by combining the co-culture with multicolor, high-throughput FACS, we can evaluate the effect of hundreds of small molecules on multi-parametric processes including: cell survival, stemness (leukemic stem cells), and myeloid differentiation on the primary AML cells at a single-cell level. This method streamlines the identification of potential therapeutic agents, but also facilitates combinatorial screening aiming, for instance, at dissecting the regulatory pathways in a patient-specific manner.

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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Drug, FACS, High throughput, Leukemia, Screen, Stem cell
in
Bio-protocol
volume
12
issue
6
article number
e4353
publisher
Bio-protocol LLC
external identifiers
  • scopus:85128129686
  • pmid:35434186
ISSN
2331-8325
DOI
10.21769/BioProtoc.4353
language
English
LU publication?
yes
id
f2d9624d-1e72-41bf-bb9c-7a754e353b13
date added to LUP
2022-07-01 15:02:53
date last changed
2024-06-13 17:56:13
@article{f2d9624d-1e72-41bf-bb9c-7a754e353b13,
  abstract     = {{<p>Ex vivo culture of primary acute myeloid leukemia (AML) cells is notoriously difficult due to spontaneous differentiation and cell death, which hinders mechanistic and translational studies. To overcome this bottleneck, we have implemented a co-culture system, where the OP9-M2 stromal cells support the growth, but most notably limit the differentiation of primary AML cells, thus allowing for mechanistic studies in vitro. Additionally, the co-culture on OP9-M2 stromal is superior in preserving surface marker expression of primary (adult and pediatric) AML cells in comparison to stroma-free culture. Thus, by combining the co-culture with multicolor, high-throughput FACS, we can evaluate the effect of hundreds of small molecules on multi-parametric processes including: cell survival, stemness (leukemic stem cells), and myeloid differentiation on the primary AML cells at a single-cell level. This method streamlines the identification of potential therapeutic agents, but also facilitates combinatorial screening aiming, for instance, at dissecting the regulatory pathways in a patient-specific manner.</p>}},
  author       = {{Baudet, Aurélie and Hultmark, Simon and Ek, Fredrik and Magnusson, Mattias}},
  issn         = {{2331-8325}},
  keywords     = {{Drug; FACS; High throughput; Leukemia; Screen; Stem cell}},
  language     = {{eng}},
  month        = {{03}},
  number       = {{6}},
  publisher    = {{Bio-protocol LLC}},
  series       = {{Bio-protocol}},
  title        = {{Small Molecule Screening of Primary Human Acute Myeloid Leukemia Using Co-culture and Multiplexed FACS Analysis}},
  url          = {{http://dx.doi.org/10.21769/BioProtoc.4353}},
  doi          = {{10.21769/BioProtoc.4353}},
  volume       = {{12}},
  year         = {{2022}},
}